Fluorescence co-staining by Hoechst 33342 and acridine orange to determine ciliate species (phylum: Ciliophora)

The work describes a new method of co-staining with the fluorochromes Hoechst 33342 (Ho342) and acridine orange (AO). The method of co-staining is simple to use and, therefore, recommended especially to systematically determine the various species of ciliates (phylum: Ciliophora). The method specifically stains nuclei (in general: DNA) in blue color and also stains phagosomes, lysosomes, acidosomes and in certain cases trichocysts in colors ranging from green to red. Fine structures along the ciliary patterns can be shown by using AO to stain ciliary vacuoles and granules. Correct interpretation of the staining results is discussed in detail. The method allows life-cell-imaging, may be applied to determine any species of ciliates and does not suffer from artifacts. Hence co-staining with Ho342 and AO replaces supravital staining using methyl green-pyronin. The method is successfully applied to determine ciliate species during multi-year monitoring of the natural reserve “Verbrannte Maar“ / „Hellenmaar“ (SU-044) near Bornheim (Rhine), Germany.

Bauer T. Fluoreszenz-Doppelfärbung mit Hoechst 33342 und Acridinorange zur Bestimmung der Arten von Ciliaten (Phylum: Ciliophora). Mikroskopie. 2019; 1: 2-19.

DOI: 10.5414/MKX00192A

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